title: Multiplex immunoassay for serological diagnosis of Mycobacterium bovis infection in cattle.
abstract: Efforts to develop a better diagnostic assay for bovine tuberculosis have shown that the sensitivity and specificity of an assay can be improved by the use of two or more antigens. As reported here, we developed a multiplex chemiluminescent immunoassay that can simultaneously detect antibody activity to 25 antigens in a single well in a 96-well plate array format. The chemiluminescent signal is captured with a digital imaging system and analyzed with a macro program that tracks each serum for its pattern of antibody activity for Mycobacterium bovis antigens. The comparison of sera from 522 infected and 1,489 uninfected animals showed that a sensitivity of 93.1% and a specificity of 98.4% can be achieved with a combination of antigens. The assay system is rapid and can be automated for use in a centralized laboratory.
authors.affiliation: Enfer Scientific, Unit T, M7 Business Park, Newhall, Naas, County Kildare, Ireland.
authors.forename: Clare
authors.initials: C
authors.lastname: Whelan
authors.forename: Eduard
authors.initials: E
authors.lastname: Shuralev
authors.forename: Grainne
authors.initials: G
authors.lastname: O'Keeffe
authors.forename: Paula
authors.initials: P
authors.lastname: Hyland
authors.forename: Hang Fai
authors.initials: HF
authors.lastname: Kwok
authors.forename: Philip
authors.initials: P
authors.lastname: Snoddy
authors.forename: Amanda
authors.initials: A
authors.lastname: O'Brien
authors.forename: Marie
authors.initials: M
authors.lastname: Connolly
authors.forename: Padraig
authors.initials: P
authors.lastname: Quinn
authors.forename: Matt
authors.initials: M
authors.lastname: Groll
authors.forename: Todd
authors.initials: T
authors.lastname: Watterson
authors.forename: Sara
authors.initials: S
authors.lastname: Call
authors.forename: Kevin
authors.initials: K
authors.lastname: Kenny
authors.forename: Anthony
authors.initials: A
authors.lastname: Duignan
authors.forename: Mary Jo
authors.initials: MJ
authors.lastname: Hamilton
authors.forename: Bryce M
authors.initials: BM
authors.lastname: Buddle
authors.forename: James A
authors.initials: JA
authors.lastname: Johnston
authors.forename: William C
authors.initials: WC
authors.lastname: Davis
authors.forename: Shane A
authors.initials: SA
authors.lastname: Olwill
authors.forename: John
authors.initials: J
authors.lastname: Clarke
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D000942:Antigens, Bacterial / Q000276:immunology; D002417:Cattle; D007118:Immunoassay; D008163:Luminescent Measurements / Q000379:methods*; D009163:Mycobacterium bovis / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D014380:Tuberculosis, Bovine / Q000175:diagnosis*
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D011994:Recombinant Proteins